Antibody micro arrays are an emerging technology that has the potential to efficiently and quantitatively analyze many proteins in thousands of samples. Therefore, this technology can provide a badly needed mechanism for determining the clinical usefulness of individual or profiles of potential biomarkers. At present, though, antibody micro array analysis lacks the guiding principles, standard procedures, honed laboratory practices, foundational statistics and supporting software to produce quality results on a large scale. Therefore, we have started to develop both the protocols and supporting software ("ProMAT") specifically for analyzing protein micro array data. The goal of this proposal is to advance antibody micro array to the point that it can be used as a routine tool for clinical biomarker validation. To accomplish this goal, we plan to establish novel protocols for the use of internal and external standards, develop the statistical foundation for evaluating data quality at all stages of data collection and analysis, and develop a sophisticated bioinformatics tool for rapid data analysis, including advanced quality control features. This system will also aid in identifying sources of data variability and will allow for data normalization. We will use an iterative process that cycles through resolving assay requirements, deriving statistical methods and composing prototype software, honing laboratory practices and assay procedures, and evaluating progress through repetitive experimentation that focuses on assessing data quality and reproducibility. At the completion of this work, we will have uncovered guiding principles and requirements for conducting antibody micro array experiments and developed an integrated system for rapidly generating the high quality data required to evaluate the clinical potential of biomarker profiles.